Browse > Article
http://dx.doi.org/10.1186/s40824-016-0050-x

Multiphoton imaging of myogenic differentiation in gelatin-based hydrogels as tissue engineering scaffolds  

Kim, Min Jeong (Department of Cogno-Mechatronics Engineering, College of Nanoscience & Nanotechnology, Pusan National University)
Shin, Yong Cheol (Department of Cogno-Mechatronics Engineering, College of Nanoscience & Nanotechnology, Pusan National University)
Lee, Jong Ho (Department of Cogno-Mechatronics Engineering, College of Nanoscience & Nanotechnology, Pusan National University)
Jun, Seung Won (Department of Cogno-Mechatronics Engineering, College of Nanoscience & Nanotechnology, Pusan National University)
Kim, Chang-Seok (Department of Cogno-Mechatronics Engineering, College of Nanoscience & Nanotechnology, Pusan National University)
Lee, Yunki (Department of Molecular Science and Technology, Ajou University)
Park, Jong-Chul (Department of Medical Engineering, Cellbiocontrol Laboratory, Yonsei University College of Medicine)
Lee, Soo-Hong (Department of Biomedical Science, CHA University)
Park, Ki Dong (Department of Molecular Science and Technology, Ajou University)
Han, Dong-Wook (Department of Cogno-Mechatronics Engineering, College of Nanoscience & Nanotechnology, Pusan National University)
Publication Information
Biomaterials Research / v.20, no.1, 2016 , pp. 8-14 More about this Journal
Abstract
Background: Hydrogels can serve as three-dimensional (3D) scaffolds for cell culture and be readily injected into the body. Recent advances in the image technology for 3D scaffolds like hydrogels have attracted considerable attention to overcome the drawbacks of ordinary imaging technologies such as optical and fluorescence microscopy. Multiphoton microscopy (MPM) is an effective method based on the excitation of two-photons. In the present study, C2C12 myoblasts differentiated in 3D gelatin hydroxyphenylpropionic acid (GHPA) hydrogels were imaged by using a custom-built multiphoton excitation fluorescence microscopy to compare the difference in the imaging capacity between conventional microscopy and MPM. Results: The physicochemical properties of GHPA hydrogels were characterized by using scanning electron microscopy and Fourier-transform infrared spectroscopy. In addition, the cell viability and proliferation of C2C12 myoblasts cultured in the GHPA hydrogels were analyzed by using Live/Dead Cell and CCK-8 assays, respectively. It was found that C2C12 cells were well grown and normally proliferated in the hydrogels. Furthermore, the hydrogels were shown to be suitable to facilitate the myogenic differentiation of C2C12 cells incubated in differentiation media, which had been corroborated by MPM. It was very hard to get clear images from a fluorescence microscope. Conclusions: Our findings suggest that the gelatin-based hydrogels can be beneficially utilized as 3D scaffolds for skeletal muscle engineering and that MPM can be effectively applied to imaging technology for tissue regeneration.
Keywords
Hydrogel; 3D scaffolds; Multiphoton microscopy; C2C12 myoblast; Myogenic differentiation;
Citations & Related Records
Times Cited By KSCI : 3  (Citation Analysis)
연도 인용수 순위
1 Ratner BD, Hoffman AS, Schoen FJ, Lemons JE. Biomaterials science: an introduction to materials in medicine. 3rd ed. San Diego: Academic Press; 2004.
2 Hoffman AS. Hydrogels for biomedical applications. Adv Drug Delivery Rev. 2002;54:3-12.   DOI
3 Kytai TN, Jennifer LW. Photopolymerizable hydrogels for tissue engineering applications. Biomaterials. 2002;23:4307-14.   DOI
4 Serafim A, Dragusin DM, Zecheru T, Dubruel P, Petre D, Ciocan LT, et al. Gelatin hydrogels: effect of ethylene oxide based synthetic crosslinking agents on the physico-chemical properties. Dig J Nanomater Bios. 2013;8:101-10.
5 Park S, Lee KS, Bozoklu G, Cai W, Nguyen ST, Ruoff RS. Graphene oxide papers modified by divalent ions-enhancing mechanical properties via chemical cross-linking. ACS Nano. 2008;2:572-8.   DOI
6 Wang JW, Wong AM, Flores J, Vosshall LB, Axel R. Two-photon calcium imaging reveals an odore-voked map of activity in the fly brain. Cell. 2003;112:271-82.   DOI
7 Oertner TG. Functional imaging of single synapses in brain slices. Exp Physiol. 2002;87:733-6.   DOI
8 Lee HS, Lee HD, Jeong MY, Kim CS. Wavelength-swept cascaded Raman fiber laser around 1300 nm for OCT imaging. J Opt Soc Korea. 2015;19:154-8.   DOI
9 Chatterjee K, Lin-Gibson S, Wallace WE, Parekh SH, Lee YJ, Cicerone MT, et al. The effect of 3D hydrogel scaffold modulus on osteoblast differentiation and mineralization revealed by combinatorial screening. Biomaterials. 2010;31:5051-62.   DOI
10 Rose CR, Kovalchuk Y, Eilers J, Konnerth A. Two-photon $Na^+$ imaging in spines and fine dendrites of central neurons. Pflugers Arch. 1999;439:201-7.
11 Nagai Y, Yokoi H, Kaihara K, Naruse K. The mechanical stimulation of cells in 3D culture within a self-assembling peptide hydrogel. Biomaterials. 2012;33:1044-51.   DOI
12 Wang LS, Du C, Chung JE, Kurikawa M. Enzymatically cross-linked gelatin-phenol hydrogels with a broader stiffness range for osteogenic differentiation of human mesenchymal stem cells. Acta Biomater. 2012;8:1826-37.   DOI
13 Sahu A, Choi WI, Tae G. A stimuli-sensitive injectable graphene oxide composite hydrogel. Chem Commun. 2012;48:5801-940.   DOI
14 Denk W, Strickler JH, Webb WW. Two-photon laser scanning fluorescence microscopy. Science. 1990;248:73-6.   DOI
15 Jeon BH, Chae YG, Hwang SS, Kim DK, Oak C, Park EK, et al. Multimodal imaging of sarcopeni using optical coherence tomography and ultrasound in rat model. J Opt Soc Korea. 2014;18:55-9.   DOI
16 Conchello JA, Lichtman JW. Optical sectioning microscopy. Nat Methods. 2005;2:920-31.   DOI
17 Weber M, Huisken J. Light sheet microscopy for real-time developmental biology. Curr Opin Genet Dev. 2011;21:566-72.   DOI
18 Gao L, Shao L, Higgins CD, Poulton JS, Peifer M, Davidson MW, et al. Noninvasive imaging beyond the diffraction limit of 3D dynamics in thickly fluorescent specimens. Cell. 2012;151:1370-85.   DOI
19 Wicker K, Heintzmann R. Interferometric resolution improvement for confocal microscopes. Opt Express. 2007;15:12206-16.   DOI
20 Kobat D, Horton NG, Xu C. In vivo two-photon microscopy to 1.6-mm depth in mouse cortex. J Biomed Opt. 2011;16:106014.   DOI
21 Richard KPB, David WP. Two-photon excitation microscopy for the study of living cells and tissues. Cell Biol. 2014;4:1124-59.
22 Ammasi P, Paul S, Colten N, Raymond K. An evaluation of two-photon excitation versus confocal and digital deconvolution fluorescence microscopy imaging in xenopus morphogenesis. Microsc Res Techniq. 1999;47:172-81.   DOI
23 Karel S, Ryohei Y. Principle of two-photon excitation microscopy and its applications to neuroscience. Neuron. 2006;50:823-39.   DOI
24 Majewska A, Yiu G, Yuste R. A custom-made two-photon microscope and deconvolution system. Pflugers Arch. 2000;441:398-408.   DOI
25 Thorling CA, Crawford D, Burczynski FJ, Liu X, Liau I, Roberts MS. Multiphoton microscopy in defining liver function. J Biomed Opt. 2014;19:90901.   DOI
26 Park KM, Jun I, Joung YK, Shin H, Park KD. In situ hydrogelation and RGD conjugation of tyramine-conjugated 4-arm PPO-PEO block copolymer for injectable bio-mimetic scaffolds. Soft Matter. 2011;7:986-92.   DOI
27 Shin YC, Lee JH, Jin L, Kim MJ, Kim YJ, Hyun JK, et al. Stimulated myoblast differentiation on graphene oxide-impregnated PLGA-collagen hybrid fibre matrices. J Nanobiotechnol. 2015;13:21.   DOI
28 Yang T, Liu L, Liu J, Chen M, Wang J. Cyanobacterium metallothionein decorated graphene oxide nanosheets for highly selective adsorption of ultra-trace cadmium. J Mater Chem. 2012;22:21909-16.   DOI
29 Liu Z, Jiang L, Galli F, Nederlof I, Olsthoorn RCL, Lamers GEM, et al. A graphene oxide streptavidin complex for biorecognition - towards affinity purification. Adv Funct Mater. 2010;20:2857-65.   DOI
30 Lee Y, Bae JW, Oh DH, Park KM, Chun YW, Sung HJ, et al. In situ forming gelatin-based tissue adhesives and their phenolic content-driven properties. J Mater Chem B. 2013;1:2407-14.   DOI
31 Lee Y, Bae JW, Lee JW, Suh W, Park KD. Enzyme-catalyzed in situ forming gelatin hydrogels as bioactive wound dressings: effects of fibroblast delivery on wound healing efficacy. J Mater Chem B. 2014;2:7712-8.   DOI
32 Wang LS, Boulaire J, Chan PPY, Chung JE, Kurikawa M. The role of stiffness of gelatin-hydroxyphenylpropionic acid hydrogels formed by enzyme-mediated crosslinking on the differentiation of human mesenchymal stem cell. Biomateirals. 2010;31:8608-16.   DOI
33 Neffe AT, Loebus A, Zaupa A, Stoetzel C, Muller FA, Lendlein A. Gelatin functionalization with tyrosine derived moieties to increase the interaction with hydroxyapatite fillers. Acta Biomater. 2011;7:1693-701.   DOI
34 Liu XH, Ma PX. Phase separation, pore structure, and properties of nanofibrous gelatin scaffolds. Biomaterials. 2009;30:4094-103.   DOI
35 Silva SS, Mano JF, Reis RL. Potential applications of natural origin polymerbased systems in soft tissue regeneration. Crit Rev Biotechnol. 2010;30:200-21.   DOI
36 Huang S, Fu XB. Naturally derived materials-based cell and drug delivery systems in skin regeneration. J Control Release. 2010;142:149-59.   DOI
37 Yuan SJ, Xiong G, Roguin A, Choong C. Immobilization of gelatin onto poly(glycidyl methacrylate)-grafted polycaprolactone substrates for improved cell-material interactions. Biointerphases. 2012;7:30.   DOI
38 Shin YC, Lee JH, Jin OS, Lee EJ, Jin L, Kim C, et al. RGD peptide-displaying M13 bacteriophage/PLGA nanofibers as cell-adhesive matrices for smooth muscle cells. J Korean Phys Soc. 2015;66:12-6.   DOI
39 Shin YC, Lee JH, Jin L, Kim MJ, Kim C, Hong SW, et al. Cell-adhesive matrices composed of RGD peptide-displaying M13 bacteriophage/poly(lactic-co-glycolic acid) nanofibers beneficial to myoblast differentiation. J Nanosci Nanotechnol. 2015;15:7907-12.   DOI
40 Shin YC, Lee JH, Kim MJ, Hong SW, Oh JW, Kim CS, et al. Stimulating effect of graphene oxide on myogenesis of C2C12 myoblasts on PLGA/RGD nanofiber matrices. J Biol Eng. 2015;9:22.   DOI
41 Shin YC, Jin L, Lee JH, Jun S, Hong SW, Kim CS, et al. Graphene oxideincorporated PLGA-collagen fibrous matrices as biomimetic scaffolds for vascular smooth muscle cells. Sci Adv Mater. 2015;7:1-6.   DOI